Analysis of tissue-substrate adhesion by hyperspectral surface plasmon resonance microscopy.

Abstract

The preparation of histology slides is a critical step in histopathology, and poor-quality histology slides with weak adhesion of tissue sections to the substrate often affect diagnostic accuracy and sometimes lead to diagnostic failure due to tissue section detachment. This issue has been of concern and some methods have been proposed to enhance tissue-substrate adhesion. Unfortunately, quantitative analysis of the adhesion between tissue sections and glass slides is still challenging. In this work, the adhesion of mouse brain tissue sections on gold-coated glass slides was analyzed using a laboratory-fabricated hyperspectral surface plasmon resonance microscopy (HSPRM) system that enabled single-pixel spectral SPR sensing and provided two-dimensional (2D) distribution of resonance wavelengths (RWs). The existence of the nanoscale water gap between the tissue section and the substrate was verified by fitting the RW measured in each pixel using the five-layer Fresnel reflection model. In addition, a 2D image of the tissue-substrate adhesion distance (AD) was obtained from the measured 2D distribution of RWs. The results showed that tissue-substrate AD was 20-35 nm in deionized water and 4-24 nm in saline solution. The HSPRM system used in this work has a wide wavelength range of 400-1000 nm and can perform highly sensitive and label-free detection over a large dynamic detection range with high spectral and spatial resolutions, showing significant potential applications in stain-free tissue imaging, quantitative analysis of tissue-substrate adhesion, accurate identification of tumor cells, and rapid histopathological diagnosis.